目的 建立了HPLC-ESI-MS/MS法同时测定四神丸中五味子甲素、五味子乙素、五味子醇甲、五味子醇乙、五味子酯甲、补骨脂素、异补骨脂素、吴茱萸碱和吴茱萸次碱9种有效成分的分析方法。方法 HPLC采用色谱柱Agilent Zorbax Eclipse Plus C₁₈柱（100 mm×4.6 mm，3.5 μm），流动相为甲醇-0.1%甲酸水溶液，梯度洗脱，体积流量0.4 mL/min，进样量20 μL；质谱采用电喷雾离子源、正离子模式（ESI⁺），通过多反应监测（MRM）同时对四神丸中的9种有效成分进行定量分析。结果 四神丸中9种有效成分五味子甲素、五味子乙素、五味子醇甲、五味子醇乙、五味子酯甲、补骨脂素、异补骨脂素、吴茱萸碱和吴茱萸次碱的线性范围分别为8.50～850.00 ng/mL（r=0.999 7）、1.32～132.00 ng/mL（r=0.997 4）、9.60～960.00 ng/mL（r=0.999 8）、12.00～1200.00 ng/mL（r=0.999 3）、11.50～1 150.00 ng/mL（r=0.997 9）、21.70～2 170.00 ng/mL（r=0.999 7）、23.80～2 380.00 ng/mL（r=0.999 6）、10.70～1 070.00 ng/mL（r=0.999 5）、8.54～854.00 ng/mL（r=0.998 0），平均加样回收率分别为98.3%、100.3%、99.2%、100.4%、99.1%、97.7%、99.0%、98.9%、100.3%，RSD值分别为2.21%、1.78%、2.19%、2.23%、2.18%、3.03%、2.51%、2.72%、2.10%。8批四神丸样品中9种有效成分的质量分数分别为67.6～425.6、0～131.5、2.1～258.0、0～71.2、23.2～678.8、806.4～1 310.8、718.5～1293.7、11.5～123.2、10.9～62.4 μg/g。结论 所建立的分析方法简单、灵敏度高、专属性好，可应用于不同厂家不同生产批次的四神丸中有效成分的测定及质量控制。

Objective To develop and validate an high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of nine major bioactive components (deoxyschizandrin, γ-schizandrin, schizandrin, schizandrol B, schisantherin A, psoralen, isopsoralen, evodiamine, and rutaecarpine) in Sishen Pills.Methods The chromatographic separation was performed on an Agilent Zorbax Eclipse Plus C₁₈ column (100 mm×4.6mm, 3.5 μm) with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.4 mL/min, and the injection volume was 20 μL.The nine major bioactive components were detected using an electrospray ionization source in positive ionization mode (ESI⁺) and quantified by multiple reaction monitor (MRM) scanning at the same time.Results The linear ranges of deoxyschizandrin, γ-schizandrin, schizandrin, schizandrol B, schisantherin A, psoralen, isopsoralen, evodiamine, and rutaecarpine were 8.50-850.00 ng/mL (r=0.999 7), 1.32-132.00 ng/mL (r=0.997 4), 9.60-960.00 ng/mL (r=0.999 8), 12.00-1 200.00 ng/mL (r=0.999 3), 11.50-1 150.00 ng/mL (r=0.997 9), 21.70-2 170.00 ng/mL (r=0.999 7), 23.80-2 380.00 ng/mL (r=0.999 6), 10.70-1 070.00 ng/mL (r=0.999 5), 8.54-854.00 ng/mL (r=0.998 0), and the average recoveries were 98.3%(RSD=2.21%), 100.3%(RSD=1.78%), 99.2%(RSD=2.19%), 100.4%(RSD=2.23%), 99.1%(RSD=2.18%), 97.7%(RSD=3.03%), 99.0%(RSD=2.51%), 98.9%(RSD=2.72%), and 100.3%(RSD=2.10%), respectively.The contents of eight batches of the nine major bioactive components were 67.6-425.6, 0-131.5, 2.1-258.0, 0-71.2, 23.2-678.8, 806.4-1310.8, 718.5-1293.7, 11.5-123.2, and 10.9-62.4 μg/g, respectively.Conclusion The developed method is simple, specific, and sensitive, and it can be applied for the determination of nine major bioactive components and the quality control of Sishen Pills collected from different production batches.

山西省自然科学基金资助项目（201701D121145）